Characterization of Envelopment Process of viral Nucleocapsids in Herpes Simplex Virus-Infected Cells

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We have quantitatively studied an intracellular multiplication of herpes simplex virus type 1 (HSV-1) in Vero cells. Characterization of the inhibition of the multiplication of HSV-1 by ammonium chloride revealed that the formation of the virus particles (i, e., the envelopment of viral nucleocapsids) takes place in the intracellular acidic compartments and that the acidity in the compartments is essential for the envelopment. To examine the effect of the agent on the synthesis and transport of spike proteins of virus, the appearance of ability to adsorb the sensitized erythrocytes on the HSV-1-infected cell surface was characterized. The kinetics of appearance indicated that both synthesis and transport of spike proteins are not affected by the agent.Since the most acidic and the most agent-sensitive compartment among intracellular organelles is known to be the Golgi complex, we examined the role of the Golgi complex in the envelopment as well as in the formation of infectious progeny virus in the HSV-1-infected cells. Since Orefeldin A (BFA) is recently reported to cause disassembly of the Golgi complex, we characterized the effect of BFA on the HSV-1 multiplication, showing that (1) there are two different (i. e., agent-sensitive and -insensitive) pathways for the formation of virus particles in the HSV-1-infected cells and that (2) the infectious progeny virus arises exclusively from the agent-sensitive pathway.These results indicated the crucial role of the Golgi complex in the envelopment of HSV-1 that lead to the formation of the infectious progeny virus.